TY - GEN
T1 - Ambiguities and completeness of SAS data analysis
T2 - 3rd International School and Workshop on Complex and Magnetic Soft Matter Systems: Structure and Physico - Mechanical Properties, CMSMS 2017
AU - Zabelskii, D.V.
AU - Vlasov, A.V.
AU - Ryzhykau, Yu L.
AU - Murugova, T.N.
AU - Brennich, M.
AU - Soloviov, D.V.
AU - Ivankov, O.I.
AU - Borshchevskiy, V.I.
AU - Mishin, A.V.
AU - Rogachev, A.V.
AU - Round, A.
AU - Dencher, N.A.
AU - Büldt, G.
AU - Gordeliy, V.I.
AU - Kuklin, A.I.
PY - 2018/3/28
Y1 - 2018/3/28
N2 - The method of small angle scattering (SAS) is widely used in the field of biophysical research of proteins in aqueous solutions. Obtaining low-resolution structure of proteins is still a highly valuable method despite the advances in high-resolution methods such as X-ray diffraction, cryo-EM etc. SAS offers the unique possibility to obtain structural information under conditions close to those of functional assays, i.e. in solution, without different additives, in the mg/mL concentration range. SAS method has a long history, but there are still many uncertainties related to data treatment. We compared 1D SAS profiles of apoferritin obtained by X-ray diffraction (XRD) and SAS methods. It is shown that SAS curves for X-ray diffraction crystallographic structure of apoferritin differ more significantly than it might be expected due to the resolution of the SAS instrument. Extrapolation to infinite dilution (EID) method does not sufficiently exclude dimerization and oligomerization effects and therefore could not guarantee total absence of dimers account in the final SAS curve. In this study, we show that EID SAXS, EID SANS and SEC-SAXS methods give complementary results and when they are used all together, it allows obtaining the most accurate results and high confidence from SAS data analysis of proteins.
AB - The method of small angle scattering (SAS) is widely used in the field of biophysical research of proteins in aqueous solutions. Obtaining low-resolution structure of proteins is still a highly valuable method despite the advances in high-resolution methods such as X-ray diffraction, cryo-EM etc. SAS offers the unique possibility to obtain structural information under conditions close to those of functional assays, i.e. in solution, without different additives, in the mg/mL concentration range. SAS method has a long history, but there are still many uncertainties related to data treatment. We compared 1D SAS profiles of apoferritin obtained by X-ray diffraction (XRD) and SAS methods. It is shown that SAS curves for X-ray diffraction crystallographic structure of apoferritin differ more significantly than it might be expected due to the resolution of the SAS instrument. Extrapolation to infinite dilution (EID) method does not sufficiently exclude dimerization and oligomerization effects and therefore could not guarantee total absence of dimers account in the final SAS curve. In this study, we show that EID SAXS, EID SANS and SEC-SAXS methods give complementary results and when they are used all together, it allows obtaining the most accurate results and high confidence from SAS data analysis of proteins.
UR - http://www.scopus.com/inward/record.url?scp=85045912396&partnerID=8YFLogxK
U2 - 10.1088/1742-6596/994/1/012017
DO - 10.1088/1742-6596/994/1/012017
M3 - Conference contribution
VL - 994
T3 - Journal of Physics: Conference Series
BT - 3rd International Summer School and Workshop "Complex and Magnetic Soft Matter Systems: Physico-Mechanical Properties and Structure"
A2 - Raikher, Y.
A2 - Rogachev, A.
A2 - Bunoiu, M.
A2 - Balasoiu, M.
PB - Institute of Physics Publishing
Y2 - 28 June 2017 through 30 June 2017
ER -